Advertisement

Translational Stroke Research

, Volume 10, Issue 1, pp 57–66 | Cite as

Evaluation of the Neuroprotective Effect of Sirt3 in Experimental Stroke

  • Rajkumar VermaEmail author
  • Rodney M. Ritzel
  • Joshua Crapser
  • Brett D. Friedler
  • Louise D. McCullough
Original Article

Abstract

Sirtuins (Sirt) are a family of NAD+ dependent histone deacetylase (HDAC) proteins implicated in aging, cell cycle regulation, and metabolism. These proteins are involved in the epigenetic modification of neuromodulatory proteins after strokevia acetylation/deacetylation. The specific role of Sirt3, a mitochondrial sirtuin, in post-stroke injury has been relatively unexplored. Using male Sirt3 knockout (KO) mice and wild-type littermates (WT), we show that Sirt3 KO mice show significant neuroprotection at 3 days after ischemia/reperfusion (I/R) or stroke injury. The deacetylation activity of Sirt3, measured as the amount of reduced acetylated lysine, was increased after stroke. Stroke-induced increases in liver kinase 1 (LKB1) activity were also reduced in KO mice at 3 days after stroke. On further investigation, we found that the levels of Sirt1, another important member of the Sirtuin family, were increased in the brains of Sirt3 KO mice after stroke. To determine the translational relevance of these findings, we then tested the effects of pharmacological inhibition of Sirt3. We found no benefit of Sirt3 inhibition despite clear evidence of deacetylation. Overall, these data suggest that Sirt3 KO mice show neuroprotection by a compensatory rise in Sirt1 rather than the loss of Sirt3 after stroke. Further analysis reveals that the beneficial effects of Sirt1 might be mediated by a decrease in LKB1 activity after stroke. Finally, our data clearly demonstrate the importance of using both pharmacological and genetic methods in pre-clinical stroke studies.

Keywords

Sirtuins Stroke Neuroprotection Deacetylase LKB1 

Notes

Funding

This work was supported by National Institutes of Health grants R01NSO77769 (to Louise D McCullough) and an AHA postdoctoral fellowship 14POST20380612 (to Rajkumar Verma).

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Ethical Approval

All animal protocols were approved by the University’s Institutional Animal Care and Use Committee at UConn Health, Farmington, CT, and were performed in accordance with National Institutes of Health guidelines.

Supplementary material

12975_2017_603_MOESM1_ESM.docx (12.8 mb)
ESM 1 (DOCX 13128 kb)

References

  1. 1.
    Writing Group M, Mozaffarian D, Benjamin EJ, Go AS, Arnett DK, Blaha MJ, et al. Heart Disease and Stroke Statistics-2016 update: a report from the American Heart Association. Circulation. 2016;133(4):e38–360.  https://doi.org/10.1161/CIR.0000000000000350.CrossRefGoogle Scholar
  2. 2.
    Robbins NM, Swanson RA. Opposing effects of glucose on stroke and reperfusion injury: acidosis, oxidative stress, and energy metabolism. Stroke. 2014;45(6):1881–6.  https://doi.org/10.1161/STROKEAHA.114.004889.CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Lo EH. A new penumbra: transitioning from injury into repair after stroke. Nat Med. 2008;14(5):497–500.  https://doi.org/10.1038/nm1735.CrossRefPubMedGoogle Scholar
  4. 4.
    Schweizer S, Meisel A, Marschenz S. Epigenetic mechanisms in cerebral ischemia. J Cereb Blood Flow Metab. 2013;33(9):1335–46.  https://doi.org/10.1038/jcbfm.2013.93.CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Gibson CL, Murphy SP. Benefits of histone deacetylase inhibitors for acute brain injury: a systematic review of animal studies. J Neurochem. 2010;115(4):806–13.  https://doi.org/10.1111/j.1471-4159.2010.06993.x.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Baltan S, Morrison RS, Murphy SP. Novel protective effects of histone deacetylase inhibition on stroke and white matter ischemic injury. Neurotherapeutics. 2013;10(4):798–807.  https://doi.org/10.1007/s13311-013-0201-x.CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Kim HJ, Rowe M, Ren M, Hong JS, Chen PS, Chuang DM. Histone deacetylase inhibitors exhibit anti-inflammatory and neuroprotective effects in a rat permanent ischemic model of stroke: multiple mechanisms of action. J Pharmacol Exp Ther. 2007;321(3):892–901.  https://doi.org/10.1124/jpet.107.120188.CrossRefPubMedGoogle Scholar
  8. 8.
    Bassett SA, Barnett MP. The role of dietary histone deacetylases (HDACs) inhibitors in health and disease. Nutrients. 2014;6(10):4273–301.  https://doi.org/10.3390/nu6104273.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Lu X, Bennet B, Mu E, Rabinowitz J, Kang Y. Metabolomic changes accompanying transformation and acquisition of metastatic potential in a syngeneic mouse mammary tumor model. J Biol Chem. 2010;285(13):9317–21.  https://doi.org/10.1074/jbc.C110.104448.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Finley LW, Carracedo A, Lee J, Souza A, Egia A, Zhang J, et al. SIRT3 opposes reprogramming of cancer cell metabolism through HIF1alpha destabilization. Cancer Cell. 2011;19(3):416–28.  https://doi.org/10.1016/j.ccr.2011.02.014.CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Nogueiras R, Habegger KM, Chaudhary N, Finan B, Banks AS, Dietrich MO, et al. Sirtuin 1 and sirtuin 3: physiological modulators of metabolism. Physiol Rev. 2012;92(3):1479–514.  https://doi.org/10.1152/physrev.00022.2011.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Scher MB, Vaquero A, Reinberg D. SirT3 is a nuclear NAD+-dependent histone deacetylase that translocates to the mitochondria upon cellular stress. Genes Dev. 2007;21(8):920–8.  https://doi.org/10.1101/gad.1527307.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Schwer B, Bunkenborg J, Verdin RO, Andersen JS, Verdin E. Reversible lysine acetylation controls the activity of the mitochondrial enzyme acetyl-CoA synthetase 2. Proc Natl Acad Sci U S A. 2006;103(27):10224–9.  https://doi.org/10.1073/pnas.0603968103.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Bai Y, Zhou T, Fu H, Sun H, Huang B. 14-3-3 interacts with LKB1 via recognizing phosphorylated threonine 336 residue and suppresses LKB1 kinase function. FEBS Lett. 2012;586(8):1111–9.  https://doi.org/10.1016/j.febslet.2012.03.018.CrossRefPubMedGoogle Scholar
  15. 15.
    Lan F, Cacicedo JM, Ruderman N, Ido Y. SIRT1 modulation of the acetylation status, cytosolic localization, and activity of LKB1. Possible role in AMP-activated protein kinase activation. J Biol Chem. 2008;283(41):27628–35.  https://doi.org/10.1074/jbc.M805711200.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Pillai VB, Sundaresan NR, Kim G, Gupta M, Rajamohan SB, Pillai JB, et al. Exogenous NAD blocks cardiac hypertrophic response via activation of the SIRT3-LKB1-AMP-activated kinase pathway. J Biol Chem. 2010;285(5):3133–44.  https://doi.org/10.1074/jbc.M109.077271.CrossRefPubMedGoogle Scholar
  17. 17.
    Khan M, Dhammu TS, Matsuda F, Singh AK, Singh I. Blocking a vicious cycle nNOS/peroxynitrite/AMPK by S-nitrosoglutathione: implication for stroke therapy. BMC Neurosci. 2015;16(1):42.  https://doi.org/10.1186/s12868-015-0179-x.CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Verma R, Friedler BD, Harris NM, McCullough LD. Pair housing reverses post-stroke depressive behavior in mice. Behav Brain Res. 2014;269:155–63.  https://doi.org/10.1016/j.bbr.2014.04.044.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Longa EZ, Weinstein PR, Carlson S, Cummins R. Reversible middle cerebral artery occlusion without craniectomy in rats. Stroke. 1989;20(1):84–91.  https://doi.org/10.1161/01.STR.20.1.84.CrossRefPubMedGoogle Scholar
  20. 20.
    Verma R, Harris NM, Friedler BD, Crapser J, Patel AR, Venna V, et al. Reversal of the detrimental effects of post-stroke social isolation by pair-housing is mediated by activation of BDNF-MAPK/ERK in aged mice. Sci Rep. 2016;6(1):25176.  https://doi.org/10.1038/srep25176.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Huang W, Huang Y, Huang RQ, Huang CG, Wang WH, Gu JM, et al. SIRT3 expression decreases with reactive oxygen species generation in rat cortical neurons during early brain injury induced by experimental subarachnoid hemorrhage. Biomed Res Int. 2016;2016:8263926–9.  https://doi.org/10.1155/2016/8263926.CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Michan S, Sinclair D. Sirtuins in mammals: insights into their biological function. Biochem J. 2007;404(1):1–13.  https://doi.org/10.1042/BJ20070140.CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Sakamoto J, Miura T, Shimamoto K, Horio Y. Predominant expression of Sir2alpha, an NAD-dependent histone deacetylase, in the embryonic mouse heart and brain. FEBS Lett. 2004;556(1–3):281–6.  https://doi.org/10.1016/S0014-5793(03)01444-3.CrossRefPubMedGoogle Scholar
  24. 24.
    Weir HJ, Murray TK, Kehoe PG, Love S, Verdin EM, O'Neill MJ, et al. CNS SIRT3 expression is altered by reactive oxygen species and in Alzheimer’s disease. PLoS One. 2012;7(11):e48225.  https://doi.org/10.1371/journal.pone.0048225.CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Woods A, Johnstone SR, Dickerson K, Leiper FC, Fryer LG, Neumann D, et al. LKB1 is the upstream kinase in the AMP-activated protein kinase cascade. Curr Biol. 2003;13(22):2004–8.  https://doi.org/10.1016/j.cub.2003.10.031.CrossRefPubMedGoogle Scholar
  26. 26.
    Kendrick AA, Choudhury M, Rahman SM, McCurdy CE, Friederich M, Van Hove JL, et al. Fatty liver is associated with reduced SIRT3 activity and mitochondrial protein hyperacetylation. Biochem J. 2011;433(3):505–14.  https://doi.org/10.1042/BJ20100791.CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Lombard DB, Alt FW, Cheng HL, Bunkenborg J, Streeper RS, Mostoslavsky R, et al. Mammalian Sir2 homolog SIRT3 regulates global mitochondrial lysine acetylation. Mol Cell Biol. 2007;27(24):8807–14.  https://doi.org/10.1128/MCB.01636-07.CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Drazic A, Myklebust LM, Ree R, Arnesen T. The world of protein acetylation. Biochim Biophys Acta. 2016;1864(10):1372–401.  https://doi.org/10.1016/j.bbapap.2016.06.007.CrossRefPubMedGoogle Scholar
  29. 29.
    Novgorodov SA, Riley CL, Keffler JA, Yu J, Kindy MS, Macklin WB, et al. SIRT3 deacetylates ceramide synthases: implications for mitochondrial dysfunction and brain injury. J Biol Chem. 2016;291(4):1957–73.  https://doi.org/10.1074/jbc.M115.668228.CrossRefPubMedGoogle Scholar
  30. 30.
    Kupis W, Palyga J, Tomal E, Niewiadomska E. The role of sirtuins in cellular homeostasis. J Physiol Biochem. 2016;72(3):371–80.  https://doi.org/10.1007/s13105-016-0492-6.CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Pillai VB, Sundaresan NR, Jeevanandam V, Gupta MP. Mitochondrial SIRT3 and heart disease. Cardiovasc Res. 2010;88(2):250–6.  https://doi.org/10.1093/cvr/cvq250.CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    McCullough LD, Zeng Z, Li H, Landree LE, McFadden J, Ronnett GV. Pharmacological inhibition of AMP-activated protein kinase provides neuroprotection in stroke. J Biol Chem. 2005;280(21):20493–502.  https://doi.org/10.1074/jbc.M409985200.CrossRefPubMedGoogle Scholar
  33. 33.
    Li J, Zeng Z, Viollet B, Ronnett GV, McCullough LD. Neuroprotective effects of adenosine monophosphate-activated protein kinase inhibition and gene deletion in stroke. Stroke. 2007;38(11):2992–9.  https://doi.org/10.1161/STROKEAHA.107.490904.CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Wang P, Xu TY, Guan YF, Tian WW, Viollet B, Rui YC, et al. Nicotinamide phosphoribosyltransferase protects against ischemic stroke through SIRT1-dependent adenosine monophosphate-activated kinase pathway. Ann Neurol. 2011;69(2):360–74.  https://doi.org/10.1002/ana.22236.CrossRefPubMedGoogle Scholar
  35. 35.
    Jackson JG, Pereira-Smith OM. Primary and compensatory roles for RB family members at cell cycle gene promoters that are deacetylated and downregulated in doxorubicin-induced senescence of breast cancer cells. Mol Cell Biol. 2006;26(7):2501–10.  https://doi.org/10.1128/MCB.26.7.2501-2510.2006.CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Bhattacharjee P, Paul S, Banerjee M, Patra D, Banerjee P, Ghoshal N, et al. Functional compensation of glutathione S-transferase M1 (GSTM1) null by another GST superfamily member, GSTM2. Sci Rep. 2013;3(1):2704.  https://doi.org/10.1038/srep02704.CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Kreiner G. Compensatory mechanisms in genetic models of neurodegeneration: are the mice better than humans? Front Cell Neurosci. 2015;9:56.  https://doi.org/10.3389/fncel.2015.00056.CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
    Kim S, Titcombe RF, Zhang H, Khatri L, Girma HK, Hofmann F, et al. Network compensation of cyclic GMP-dependent protein kinase II knockout in the hippocampus by Ca2+-permeable AMPA receptors. Proc Natl Acad Sci U S A. 2015;112(10):3122–7.  https://doi.org/10.1073/pnas.1417498112.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of NeurosciencesUConn HealthFarmingtonUSA
  2. 2.Department of NeurologyUniversity of Texas Health Science CenterHoustonUSA

Personalised recommendations